江苏农业学报
江囌農業學報
강소농업학보
Jiangsu Journal of Agricultural Sciences
2015年
5期
951-956
,共6页
郑佳%丁丹%梁彦丽%张亚东%王才林
鄭佳%丁丹%樑彥麗%張亞東%王纔林
정가%정단%량언려%장아동%왕재림
水稻%GS5%克隆%简单重复序列(SSR)
水稻%GS5%剋隆%簡單重複序列(SSR)
수도%GS5%극륭%간단중복서렬(SSR)
rice%GS5%cloning%simple sequence repeat (SSR)
为明确粒型控制基因GS5在2种极端粒型水稻中的序列差异和作用,本研究利用极端大粒型水稻品种TD70和小粒型水稻品种Kasalath对粒宽控制基因GS5进行了克隆和序列分析,根据序列比对结果设计分子标记,用来区分不同来源的GS5基因,并结合重组自交系的粒型信息分析该基因对粒型的调节作用。结果显示,该基因包含10个外显子和9个内含子,与TD70的GS5基因相比,Kasalath的GS5基因第1外显子上编码的第31位氨基酸由甘氨酸变成丙氨酸,同时在第36、37位缺失2个甘氨酸。以这2个氨基酸的缺失为基础,设计简单重复序列( SSR)分子标记并在2个品种及其重组自交系( RIL)中进行了验证。利用该标记可以在含有GS5-T的品种中扩增出216 bp的片段,在含有GS5-K的品种中扩增出210 bp的片段。对重组自交系的检测结果显示,含有GS5-T基因的株系有122个,含有 GS5-K基因的株系有118个。含 GS5-T基因的株系平均粒宽比含GS5-K基因的株系平均粒宽高出0.47 mm,千粒质量高出2.87 g,说明 GS5是水稻粒宽性状的主效调控基因。
為明確粒型控製基因GS5在2種極耑粒型水稻中的序列差異和作用,本研究利用極耑大粒型水稻品種TD70和小粒型水稻品種Kasalath對粒寬控製基因GS5進行瞭剋隆和序列分析,根據序列比對結果設計分子標記,用來區分不同來源的GS5基因,併結閤重組自交繫的粒型信息分析該基因對粒型的調節作用。結果顯示,該基因包含10箇外顯子和9箇內含子,與TD70的GS5基因相比,Kasalath的GS5基因第1外顯子上編碼的第31位氨基痠由甘氨痠變成丙氨痠,同時在第36、37位缺失2箇甘氨痠。以這2箇氨基痠的缺失為基礎,設計簡單重複序列( SSR)分子標記併在2箇品種及其重組自交繫( RIL)中進行瞭驗證。利用該標記可以在含有GS5-T的品種中擴增齣216 bp的片段,在含有GS5-K的品種中擴增齣210 bp的片段。對重組自交繫的檢測結果顯示,含有GS5-T基因的株繫有122箇,含有 GS5-K基因的株繫有118箇。含 GS5-T基因的株繫平均粒寬比含GS5-K基因的株繫平均粒寬高齣0.47 mm,韆粒質量高齣2.87 g,說明 GS5是水稻粒寬性狀的主效調控基因。
위명학립형공제기인GS5재2충겁단립형수도중적서렬차이화작용,본연구이용겁단대립형수도품충TD70화소립형수도품충Kasalath대립관공제기인GS5진행료극륭화서렬분석,근거서렬비대결과설계분자표기,용래구분불동래원적GS5기인,병결합중조자교계적립형신식분석해기인대립형적조절작용。결과현시,해기인포함10개외현자화9개내함자,여TD70적GS5기인상비,Kasalath적GS5기인제1외현자상편마적제31위안기산유감안산변성병안산,동시재제36、37위결실2개감안산。이저2개안기산적결실위기출,설계간단중복서렬( SSR)분자표기병재2개품충급기중조자교계( RIL)중진행료험증。이용해표기가이재함유GS5-T적품충중확증출216 bp적편단,재함유GS5-K적품충중확증출210 bp적편단。대중조자교계적검측결과현시,함유GS5-T기인적주계유122개,함유 GS5-K기인적주계유118개。함 GS5-T기인적주계평균립관비함GS5-K기인적주계평균립관고출0.47 mm,천립질량고출2.87 g,설명 GS5시수도립관성상적주효조공기인。
In order to identify the differences of sequences and role of extreme grain gene GS5 in rice, TD70 with extremely large grain and Kasalath with extremely small grain were used to clone GS5 genes. According to the se-quences differences, molecular markers were designed and the role of GS5 were analyzed through based on the grain shape of RIL. GS5 contained ten exons and nine introns. Compared with GS5 of TD70, GS5 of Kasalath showed a mutation changing the amino acid from Gly to Ala in +31 and deleted two glys in +36 and +37 at the first exon. Based on this deletion, a simple sequence repeat ( SSR) molecular marker was designed and veri-fied in the recombinant inbred lines ( RILs) . The marker could amplify a 216-bp fragment in rice with GS5-T and a 210-bp fragment in rice with GS5-K. In the 240 RILs, GS5-T genotype were 122 and GS5-k genotype were 118. The gain width of lines with GS5-T was 0. 47 mm wider than those of lines with GS5-K,and the 1 000-grain weight (kgw) of lines with GS5-T was 2. 87 g heavier than those of lines with GS5-K. These indicated that GS5 was the major gene controlling grain width.